Surface immunoglobulins of lymphocytes in chronic lymphocytic leukemia]

Using the fluorescence technique the presence of various immunoglobulin classes (IgG, IgA, IgM) was determined on the surface of peripheral blood lymphocytes obtained from healthy subjects (10 cases) and from patients with chronic lymphatic leukaemia (65 cases). In the investigations the effect of treatment and clinical course on the values of these parameters was taken into consideration. Nearly in all cases of this leukaemia the percent of lymphocytes binding immunoglobulins on their surface was raised in comparison with the lymphocytes of healthy subjects. Presence of one or more classes of immunoglobulins was demonstrated on the surface of leukaemic lymphocytes.     

Surface immunoglobulins of lymphocytes in chronic lymphocytic leukemia and disseminated lymphosarcoma

Immunofluorescent staining was used to study the incidence of heavy and light chain determinants on the surface of blood lymphocytes from nine normal individuals, fourteen patients with chronic lymphocytic leukemia (CLL), seven patients with lymphosarcoma cell leukemia (LSL), and one hairy cell leukemia. In CLL, 46% of the lymphocytes stained for the μ chain component of the IgM immunoglobulin; this finding suggests a clonal lymphocytic population. In contrast, the surface immunoglobulins in LSL showed a preponderance of both γ and μ chains. The concomitant distribution of both heavy chain molecules on the same lymphocyte was further shown by double fluorescent staining. These results support the hypothesis that CLL and LSL are distinct lymphoproliferative disorders and that they arise from two different types of lymphocytes.     

Cytoplasmic immunoglobulins in chronic B-lymphoid leukemia

Chronic lymphocytic leukemia B-cells (B-CLL cells) have surface immunoglobulin (Smlg). In addition, cytoplasmic immunoglobulins (Clg) have been reported in chronic lymphocytic leukemia B-cells and are useful for ascertaining monoclonality of the disease. In a study of 60 cases of chronic lymphocytic leukemia B-cells, surface immunoglobulin determinations alone demonstrated monoclonality in 38% of cases, versus 65% of cases with combined surface immunoglobulin and cytoplasmic immunoglobulins determinations. Membrane markers were studied using a panel of monoclonal antibodies. Thirty-six per cent of patients were positive for CD 10. No correlations were seen between immunologic markers and clinical stage.     

Some chronic lymphocytic leukemia cells bearing surface immunoglobulins share determinants with T cells.

One set of antigens is common to some chronic lymphocytic leukemia (CLL) cells bearing surface immunoglobulins (Ig) and normal T cells. Proliferating cells from thirty-eight patients were studied with four antisera recognizing normal human T but not B cells. These antisera were raised in rabbits against (a) Sezary cells, (b) blood lymphocytes from a patient with sex-linked agammaglobulinemia, (c) T lymphoblasts and (d) thymus cells. In four CLL cases, the cells expressed the receptor for sheep erythrocytes and lacked surface Ig. Cells from thirty-four CLL cases bore monoclonal surface Ig and did not bind sheep erythrocytes. Twelve out of these thirty-four cases of CLL had cells which were lysed by one or, more frequently, by the four anti-human T cell xenoantisera. By absorption experiments, one set of at least three antigens common to the cells of some of these CLL and T cells was defined. Depending on the patient, the cells can either carry one, some or all of the antigens of this set. However, it was also demonstrated by absorption that these cells lacked antigens particular to the T cell lineage, while the cells from T CLL cases carried both sets of antigens.     

Surface immunoglobulins of leukaemic cells

We have examined leukaemia cells from twenty-eight patients for the presence of surface immunoglobulins by a migration-inhibition technique. Immunoglobulins were identified on the cells of fourteen out of nineteen patients with chronic lymphatic leukaemia but were not present on cells from patients with chronic myeloid leukaemia or acute leukaemia. The results obtained with the migration technique were confirmed by identification of immunoglobulins in eluates and lysates of the leukaemia cells. Serial elution and lysis studies indicated that in some cases the immunoglobulins were produced by the leukaemia cells at a low but steady rate, possibly indicating a relationship between those cells and `B' cells. In other cases the immunoglobulins were readily removed from the cells and were present only in the initial eluates from the cells, suggesting that they had been adsorbed and might possibly be specific antibody. The immunoglobulin negative chronic lymphatic leukaemia cells may be related to `T' cells, a possibility supported by a high level of phytohaemagglutinin induced transformation in one case examined by this technique.     

Serum immunoglobulins and lymphocyte subsets in chronic lymphocytic leukemia

The concentrations of serum immunoglobulins were correlated to the stage of disease and the proportions of peripheral blood lymphocyte subsets in 25 untreated patients with B-cell chronic lymphocytic leukemia (CLL). Diminished levels of at least one serum immunoglobulin were present in 77% of all patients with CLL and 73% of patients with Stage 0 disease. The mean concentration of IgG, IgM, and IgA decreased with advancing stage of CLL. The percentages of total T, T-helper (TH), and T-suppressor (TS) cells in the peripheral blood were less in patients with CLL than in healthy persons, but the absolute concentrations of total T, TH, and TS cells were greater in patients with CLL than controls (P less than 0.02). The absolute number of B-cells (P less than 0.01) and null cells (P less than 0.001) was also increased in patients with CLL, particularly those patients in advanced stages of CLL. These findings suggest that the hypogammaglobulinemia associated with CLL first occurs during the earliest stage of disease and may be related to the alterations in the proportion of peripheral blood lymphocytes.     

Surface immunoglobulins in chronic lymphatic leukaemia, macroglobulinaemia and myelomatosis

The direct method of immunofluorescence was applied for the detection of surface immunoglobulins (Ig) on lymphocytes, obtained from bone marrow and peripheral blood, especially from patients with paraproteinaemia. The results in chronic lymphatic leukaemia (CLL), in the macroglobulinaemia of Waldenström (MW), and in multiple myeloma (MM) show three different patterns as far as the relationship between cytoplasmic and membrane Ig is concerned. In CLL a monoclonal proliferation of cells with surface Ig was found, but the percentage of cells with cytoplasmic fluorescence was normal. In MW the further differentiation in Ig-secreting cells seems to be intact. In MM the percentage of lymphocytes in the peripheral blood with membrane-bound Ig was within normal limits or decreased.     

Surface immunoglobulins and lymphocyte-specific surface antigens on leukaemic reticuloendotheliosis cells

Hairy cells from two cases of `leukaemic reticuloendotheliosis'' were studied for their enzyme content, adherence to nylon wool columns, phagocytosis, and the presence of surface immunoglobulins and lymphocyte-specific surface antigens. The cells reacted negatively for peroxidase, chloroacetate-esterase, alpha-naphthyl-acetate-esterase and naphthol-AS-acetate-esterase. They did not adhere to nylon wool columns nor did they show significant phagocytosis. Most of the hairy cells were found to be positively labelled for surface immunoglobulins of different classes; μ chain-positive hairy cells were predominant in number. In a homo-genate of isolated, washed hairy cells from the spleen of one case only IgM could be detected in significant amounts. With peroxidase-coupled anti-thymocyte IgG most of the hairy cells of both cases showed a ring-like labelling very similar to lymphocytes, whereas monocytes and granulocytes showed no labelling.The data suggest that hairy cells are closely related to lymphatic cells of the B-cell type. They exclude with some certainty the possibility that hairy cells are directly derived from cells of the myeloid system, particularly monocytes, or from classical reticulum cells of the lymphatic tissue.     

Chromosome changes in human chronic lymphocytic leukemia

Karyotypes were studied in B- and T-lymphocyte cultures from 66 patients with B-cell CLL and two patients with T-cell CLL. Thirty-one of the B-cell cases had not been treated for their disease; 35 had received radiotherapy, corticosteroids, or cytostatic drugs. Only one of the untreated patients had a clone with an abnormal karyotype. This was present in all her mitotic cells found in cultures containing lipopolysaccharide B (LPS, a B-cell mitogen) and 10% of those in cultures with pokeweed mitogen (PWM, a T- and B-cell mitogen). The karyotype of this clone was 46,XX,t(6;7),t(7;13),t(11;14). Four of the treated patients had clones with specific chromosome changes. These were 47,XY,+12 in 10% of leukoagglutinin (LA, T-cell mitogen) and protein A (PA, T- and B-cell mitogen) cultures in one case; 47,XX,+12,del(14) in 80% of LPS cultures and in all spontaneously dividing cells in another case; 46,XY,t(6;20) in all LPS cultures in another; and 46,XX,t(1;8) in all PA cultures in another. Both structural and numerical nonclonal chromosome aberrations (9%) were found in 24% of the different cultures of cells from untreated patients, and in 15% of the cells in 20% of the different cultures in the patient who had received treatment. Both patients with T-cell CLL had received treatment for their disease, and had a normal karyotype in all cultures.     

Cisplatin-induced apoptosis in human promyelocytic leukemia cells

Cis-diamminedichloroplatinum (II) cisplatin (CDDP) is an organometallic compound frequently used in anti-cancer therapy, in particular ovarian, testicular, and head and neck tumors. We found cisplatin was effective against human promyelocytic leukemia cell line HL-60, inhibiting cell cycle progression and inducing time- and concentration- dependent cell death. Presence of nuclear fragmentation, caspase-3 cleavage and annexin V positivity suggests cell death occurred by apoptosis, although DNA internucleosomal fragmentation was not detected. In addition, analysis of malondialdehyde (MDA) production and protein carbonylation indicated that cisplatin increased lipid peroxidation and oxidation of cell proteins. This occurrence was prevented by antioxidants such as N-acetylcysteine (N-aC) and glutathione (GSH), which, consistently, were also able to prevent CDDP-induced cell death. Collectively, these findings indicate that, besides growth inhibition, an increase of oxygen radicals and lipid degradation can account for a significant part of CDDP-induced apoptosis.     

Ultrastructural localization of immunoglobulins in hairy cell leukemia

Neoplastic cells from 13 cases of hairy cell leukemia were investigated for immunoglobulin production and lysozyme activity by an electron-immunoperoxidase technique. In 10 cases cytoplasmic immunoglobulins were found, but lysozyme activity was absent in all cases. Immunoglobulins were detected in the perinuclear space and endoplasmic reticulum and at the surface of hairy cells. Of the cases in which immunoglobulins were detected in hairy cells, nine were positive with IgM antiserum and one with IgG antiserum. The immunoglobulins were monoclonal in all cases; six were positive with lambda antiserum and three with kappa antiserum. The class and type of surface immunoglobulins were identical to those of cytoplasmic immunoglobulins in the hairy cells. These results support the conclusion that hairy cells are commonly derived from immunoglobulin-producing B cells at an earlier stage of differentiation than plasma cells.